1. Viruses
Only 1 NA
Small
Syntesizes its particles

2. Laboratory diagnosis of viruses Indirect dg.
Microscopy light (only big viruses –POXVIRUS)
electron
whole virus (Rotavirus) inclusions (Negri bodies – rabies)
PCR
Immunofluorescence – with cultivation (shell vial, task 4)
Isolation – animals, chicken embryo (10 days old), tissue culture
ELISA (Rotavirus)

3. Animals in Virology Mouse (HSV)
Suckling mouse babies (Coxsackie viruses, tick-born encefalitis)
Rabbit (cornea- herpes virus)

4. Chicken embryo (task 2) Inoculation of : Amnion fluid Yolc sac
CAM…chorioalantoid membrane A
Viruses used for inoculation of:
Amnion…influenza
Alantois…influenza for vaccine preparation
CAM…Poxviruses, HSV
Yolc sac…encephalitic viruses, Chlamydia, Rickettsia
A..air
Yolc sac Y
P..paper membrane
Y..Yolc sac
w..White w

5. Tissue cultures (task 3)
Used only 1x Usefull 2x – 10x
Primary cultures Diploid c.
Monkey kidney Human embryon. lungs (LEP)
Indestructible - from tumours
Cell culture (HELA cells)

6. Preparation of tissue cultures
Changes of cells (also medium colour)
MEM (medium)
+ ATB+ cells from tissue
+ Sample
Other – not visible in microscope
CPE (cytopatic efect)
I. Plaques (hollows)
II. Rounded cells
III. Syncitia
Visualization
Cultivation in Roux bottle
Hemadsorbtion adhesion of erytrocytes
Interference
1.Virus with CPE
2. Our virus
Monkey cells
fibroblasts
Determination of a virus
Cells are growing without CPE

7. Indirect virus diagnosis (Ig)
CFT (ag+ab + C + Indicator system)
no haemolysis haemolysis
(2 samples are needed) – task 5
ELISA
Immunofluorescence
Western blott
HIT (a.b. adhere on virus and stop agglutination) – possible only in hemagglutination viruses – task 6
Neutralisation + -

8. Shell vial (task 4)-rapid cultivation – in life threating
centrifugation
Remove of a cover glass, stick to under glass
+ sample (CSF)
1 d cultivation t 37 °C
TC on small glass in glass test-tube
Indirect IF (CMV)
Direct IF (Herpes 1,2)
Sample is shining
Conjugate
a.b. marked with an fluorescence colour - conjugate
Monoclonal a.b.
cover glass + Ag
under glass